N-acetyl-d-glucosamine (nag) supplemented food products and beverages

ABSTRACT

Food products and beverages which include N-acetyl-D-glucosamine (NAG) are provided, as are methods of their use and preparation. Embodiments of the supplemented food products and beverages are heated to high temperatures, such as those used in pasteurization, without significant adverse effects on taste, color, odor and/or texture.

CROSS REFERENCE TO RELATED APPLICATIONS

This claims the benefit of U.S. Provisional Application No. 60/423,119,filed Nov. 1, 2002, and is a continuation-in-part of PCT/US02/25121,filed Aug. 7, 2002, which claims priority from U.S. application Ser. No.09/924,865, filed Aug. 8, 2001, each of which is incorporated herein byreference.

FIELD

This application relates to food products and beverages that includeN-acetyl-D-glucosamine (NAG), as well as methods of making and usingNAG-supplemented food products and beverages.

BACKGROUND

Food and beverage supplements can supply consumers with the necessaryvitamins and minerals specified in the recommended daily allowances(RDA) provided by the U.S. government. Examples of suchnutritionally-balanced snack bars are disclosed in U.S. Pat. Nos.6,432,929; 6,391,864; 4,543,262; and 3,814,819. Examples of suchnutritionally-balanced beverages are disclosed in U.S. Pat. Nos.3,894,148; 4,309,417; 4,312,856; 4,322,407; 6,432,929; and 6,391,864 aswell as EP Application No. EP 0 681 434.

Dietary cartilage supplements are effective in reducing the symptoms ofosteoarthritis and joint pain. Examples of such cartilage supplementsinclude glucosamine (GLCN) hydrochloride, GLCN sulfate, chondroitinsulfate, hyaluronic acid (which is comprised of a repeating disaccharideof N-acetyl-D-glucosamine and D-glucuronic acid), and cetyl myristoleate(CM). Two commonly used cartilage supplements are GLCN hydrochloride andGLCN sulfate.

It has been disclosed and the industry has followed the belief thatexposure of GLCN to relatively high temperatures inactivated GLCN. Inattempt to overcome this limitation, U.S. Pat. No. 6,423,929 teachesthat beverages that include GLCN are prepared using a process thatrequires two separate heating steps to minimize chemical alteration ofGLCN. A juice drink base (without GLCN) is prepared using pasteurizationat about 195° F. for 42 seconds. A separate GLCN water-based solution isprepared at a temperature of below 160° F., such that the GLCN is notinactivated. The juice drink base and the GLCN solution are then mixedto form a GLCN-supplemented beverage. Processing a beverage using twodifferent solutions at two different temperatures could be relativelyexpensive and difficult to implement.

SUMMARY

Food products and beverages, for human or animal consumption, whichinclude N-acetyl-D-glucosamine (NAG), and methods for making and usingsuch food products and beverages, are disclosed herein. In particularexamples, the disclosed food products and beverages supplemented withNAG are exposed to high temperatures without significant adverse effectson taste, color, odor, and/or texture of the NAG-supplemented foodproducts and beverages. In alternative or additional examples, the NAGpresent in the disclosed food products and beverages does notsignificantly degrade when exposed to high temperature applications. Forexample, the amount of NAG present in a NAG food product or NAG beveragefollowing exposure to a high temperature is at least about 70% of anamount of NAG present in the food or beverage prior to the exposure tohigh temperature. In some examples, the NAG-supplemented food productsand beverages are consumed to treat or prevent disease, such as acartilage dysfunction, a food allergy, or a skin disorder.

Because the nitrogen in NAG is in the neutral amide, the use of NAG overother dietary cartilage supplements, such as GLCN, which is an acidicsugar, certain embodiments of the disclosed food products and beveragesprovide various additional advantages. For example, in some examples,NAG has no substantial effect on the pH of the supplemented food productor beverage. In particular examples, NAG also imparts sweetness to thesupplemented food or beverage, making it suitable as a replacement forsome or the entire sweetener in the food or beverage. In some examples,NAG does not participate in Maillard chemistry, and therefore does notcontribute undesirable color to the product.

DETAILED DESCRIPTION OF SEVERAL EMBODIMENTS Abbreviations and Terms

The following explanations of terms and methods are provided to betterdescribe the present disclosure and to guide those of ordinary skill inthe art in the practice of the present disclosure. As used herein and inthe appended claims, the singular forms “a” or “an” or “the” includeplural references unless the context clearly dictates otherwise. Forexample, reference to “a beverage” includes a plurality of suchbeverages and reference to “the food product” includes reference to oneor more food products and equivalents thereof known to those skilled inthe art, and so forth. Similarly, the word “or” is intended to include“and” unless the context clearly indicates otherwise. Hence “comprisingA or B” means including A, or B, or A and B.

Unless otherwise indicated, all numbers expressing quantities ofingredients, temperatures, time periods, and so forth used in thespecification and claims are to be understood as being modified by theterm “about” whether explicitly stated or not. Accordingly, unlessindicated clearly to the contrary, the numerical parameters set forthare approximations.

Unless explained otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood to one of ordinaryskill in the art to which this disclosure belongs.

Administer: To cause a subject to receive something. As used herein,administration of the disclosed food products or beverages supplementedwith NAG is oral, for example by ingestion.

Beverage: Any drink suitable for ingestion. Includes beverages in theirliquid form, such as juice or soda, concentrates, as well as those in adry or powered form, such as a tea, instant coffee, or hot chocolatemix.

Cartilage dysfunction: A disorder in a subject that results in jointpain or decreased joint mobility, for example arthritis, such asosteoarthritis.

Cartilage supplement: An agent that reduces joint pain, increases jointmobility, reduces swelling, or stimulates joint healing in a subject. Inparticular examples, it is an agent that delays or halts the onset ofosteoarthritis. Examples include, but are not limited to: NAG,glucosamine, chondroitin sulfate, hyaluronic acid, chitin, cetylmyristoleate, essential fatty acids, MSM, SAMe, oligoglucosamine, andoligomers of NAG.

Comprises: A term that means including.

High Temperature: As used herein, refers to temperatures typically usedin heat pasteurization to significantly reduce the presence ofundesirable microorganisms, or in other thermal processing methods, suchas baking, broiling, boiling, roasting, or frying. A NAG food product orNAG beverage can be exposed to a high temperature for an amount of timeas needed to achieve a desired effect, for example to destroyobjectionable microorganisms, to cook a product, or to bring a NAGproduct or NAG beverage to a boil.

Particular, non-limiting examples of high temperatures include, but arenot limited to temperatures of at least about 160° F. (about 71° C.),such as temperatures of at least about 161° F. (about 71.5° C.), atleast about 180° F. (82° C.), at least about 194° F. (about 90° C.), atleast about 200° F. (about 94° C.), at least about 212° F. (about 100°C.), at least about 220° F. (about 104° C.), at least about 280° F.(about 138° C.), at least about 300° F. (about 149° C.), at least about325° F. (about 163° C.), at least about 350° F. (about 177° C.), atleast about 375° F. (about 191° C.), at least about 400° F. (about 204°C.), at least about 450° F. (about 232° C.), or at least about 500° F.(about 260° C.).

In particular examples, high temperatures include temperatures in therange of about 160° F. to about 500° F., such at about 160° F. to about350° F., about 160° F. to about 212° F., about 160° F. to about 200° F.,or about 194° F. to about 212° F.

N-acetyl-D-glucosamine (NAG): As used herein, refers to monomers of NAG,as well as oligomers of NAG that have the same or similar thermaltolerance as disclosed herein. For example, NAG and NAG oligomers can beintroduced into a food or beverage, and be subsequently subjected orexposed to a high temperature, without a resulting significant adverseeffect on the taste, color, odor, or texture of the food or beveragesupplemented with NAG.

Oligomers of NAG are those having a degree of polymerization, such as apolymer of 2-6 NAG molecules. Examples of NAG oligomers include, but arenot limited to: dimers, trimers, tetramers, pentamers, and hexamers ofNAG, which have the same or similar thermal tolerance as disclosedherein.

NAG can be obtained from any suitable source. In certain examples, NAGis a NAG composition that is derived from shellfish, cartilage,bacteria, and/or fungal biomass.

In one example, NAG is derived from fungal biomass containing chitin(for example see PCT Publication WO 03/013435). A fungal biomass thatcontains chitin and glucan is typically degraded to produce NAG. Thechitin and glucan can be degraded enzymatically (such as using enzymessecreted by eukaryotic or prokaryotic microorganisms, for examplechitinases, glucanases, and β-N-acetyl-gluosaminidases) or chemically.When enzymes are used, the degradation reaction can be maintained at apH of from about 4.0 to about 6.0 at about 20° C. to about 45° C.

Suitable starting materials include microbial fungal sources, such asfungal sources derived from Aspergillus sp., Penicillium sp., Mucor sp.,and combinations thereof. When NAG is derived from fungal biomass, itwill not pose a hazard to persons who have shellfish allergies becausetropomyosin and other such muscle-derived proteins are not present infungal biomass. Therefore, food products and beverages containing NAGderived from fungal biomass will be tolerated by persons who haveshellfish allergies. In addition, because NAG derived from fungalbiomass is not derived from shellfish (or any animal source), suchNAG-containing food products and beverages are qualified for kosherstatus and may be consumed by strict vegetarians.

In another example, NAG is derived from a bacterial source (for exampleU.S. Patent Application No. 2002/0160459). In one embodiment, bacteria,such as E. coli, are transformed with a recombinant nucleic acidencoding N-glucosamine-6-phosphate synthase, allowing the bacteria toproduce the recombinant protein, then recovering NAG from thefermentation medium.

NAG beverage: A beverage that contains NAG, for example at least about 1mg NAG per serving, such as at least about 100 mg, at least about 1 g,or at least about 5 g NAG per serving. In particular examples, theamount of NAG in a beverage is about 250 mg to about 750 mg per serving.A heat pasteurized NAG beverage is one that includes NAG in the beveragewhen the beverage is exposed to high temperatures used in heatpasteurization.

NAG food product: A food product that contains NAG, for example at leastabout 1 mg NAG per serving, at least about 100 mg, at least about 1 g,or at least about 5 g NAG per serving. In particular examples, theamount of NAG in a food product is about 250 mg to about 750 mg perserving. A pasteurized NAG food product is one that includes NAG in thefood product when the food product is exposed to high temperatures, suchas those used in baking.

Non-acidified food product: A food product that does not contain organicacids such as citric, acetic, or fumeric.

Pasteurize: A method used to significantly reduce the presence ofobjectionable organisms (such as bacteria) in a NAG food product or NAGbeverage by exposing the food or beverage to heat or irradiation for aperiod of time. Exemplary methods of pasteurization include thermalprocessing at a high temperature (referred to herein as heatpasteurization), filtration, such as microfiltration, and irradiationprocessing (such as heating in a microwave or its industrialequivalent). Ideally, pasteurization does not substantially chemicallyalter a NAG food product or NAG beverage, and does not substantiallyaffect the taste or mouthfeel of the NAG food product or NAG beverage.As used herein, “heat pasteurization” or “heat pasteurized” does notinclude pasteurization by filtration, or irradiation.

In particular examples, heat pasteurization reduces the number of colonyforming units (cfus) present in a NAG food product or NAG beverage priorto heat pasteurization by at least 50%, such as at least 70%, at least75%, at least 80%, at least 90%, at least 95%, or even at least 98%.Heat pasteurized products can be subsequently cooled quickly to about38° F. to retard the growth of surviving organisms.

Particular, non-limiting examples of heat pasteurization temperaturesinclude temperatures of at least about 160° F. (about 71° C.), such astemperatures of at least about 161° F. (about 71.5° C.), at least about180° F. (82° C.), at least 194° F. (about 90° C.), at least about 200°F. (about 94° C.), at least about 212° F. (about 100° C.), at leastabout 220° F. (about 104° C.), at least about 280° F. (about 138° C.),or at least about 300° F. (about 149° C.). In particular examples, heatpasteurization temperatures include temperatures in the range of about161° F. to about 300° F., such at about 161° F. to about 220° F., about161° F. to about 212° F., about 161° F. to about 200° F., 165° F. toabout 220° F., about 165° F. to about 212° F., about 176° F. to about220° F., or about 176° F. to about 212° F.

Particular examples of heat pasteurization temperatures and incubationtimes include, but are not limited to: at least about 15 seconds at atemperature of at least about 160° F., at least about 30 seconds at atemperature of at least about 161° F., or at least about 20 minutes atabout 350° F. Other examples include, but are not limited to: about 161°F. for 15 seconds, about 195° F. for about 42 seconds (such as about195±4° F. for about 42±4 seconds), about 200° F. for less than 40seconds (such as about 200±5° F. for about 40±5 seconds), about 165° F.for about 3 minutes (such as about 165±5° F. for about 180±10 seconds),and at or above 280° F. for about 1-2 seconds (for example toultrapasteurize milk). If ultrapasteurization is desired, pasteurizationtemperatures can be increased to about 280° F. or greater (such as about300° F.), with incubation for a shorter period of time, such as 1-2seconds.

Preventing disease: A therapeutic intervention that inhibits the fulldevelopment of a disease, for example preventing development ofosteoarthritis in a subject having cartilage dysfunction.

Serving: A serving is the amount of food or beverage a person or animalwould customarily eat in one time. The serving size can often times befound on the Nutrition Facts label on the food or beverage. Servingsizes are also shown on the USDA Food Pyramid. For bulk products, suchas breakfast cereal and flour, a serving is usually represented incommon household terms, such as cup, tablespoon, teaspoon, or fluidounce. For products that come in discrete units, such as bread andcookies, a serving size is usually listed as the number of units thatconstitute a serving, such as three cookies or two slices of bread.

Shellfish: A term for mollusks and crustaceans used as food. Exemplaryshellfish include clams, snails, mussels, oysters, scallops, shrimp,lobster, and crayfish. Components of the shell or exoskeleton of theseorganisms can be converted into GLCN using known techniques.

Shellfish protein: A protein present in a shellfish, such as those thatare allergenic in humans having shellfish allergies. Exemplary shellfishproteins include, but are not limited to, shellfish muscle proteins,such as tropomyosin.

Skin disorder: A disease or disorder in a subject that negativelyaffects the skin, and benefits from collagen formation. Examplesinclude, but are not limited to: a wound, wrinkles, and acne. When NAGis used to treat a skin disorder, NAG can be introduced into productsused on the skin, such as topical lotions and creams. Alternatively orin addition, NAG can be introduced into food products and beverages andconsumed by a subject in need of treatment or prevention of a skindisorder.

Subject: Living multicellular vertebrate organisms, a category whichincludes both human and veterinary subjects, for example, mammals,rodents, and birds.

Therapeutically Effective Amount: An amount sufficient to achieve adesired biological effect. In one example, it is an amount that iseffective to alleviate or reduce symptoms associated with cartilagedysfunction, such as pain, swelling, and decreased mobility, by morethan a desired amount. In another example, it is an amount that iseffective to stabilize symptoms associated with cartilage dysfunction,such that the symptoms do not worsen. In particular examples, it is aconcentration of NAG that is effective to alleviate, reduce, orstabilize symptoms associated with cartilage dysfunction, alone or incombination with other agent, such as in a subject to whom NAG isadministered.

In one example, it is an amount that is effective to alleviate or reducesymptoms associated with a skin disorder, such as promoting the healingof a wound or reducing the appearance of wrinkles, by more than adesired amount. In another example, it is an amount that is effective tostabilize symptoms associated with a skin disorder, such that thesymptoms do not worsen. In particular examples, it is a concentration ofNAG that is effective to alleviate, reduce, or stabilize symptomsassociated with a skin disorder, alone or in combination with otheragent, such as in a subject to whom NAG is administered.

In one example, a therapeutically effective amount also includes aquantity of NAG sufficient to achieve a desired effect in a subjectbeing treated. For instance, it can be an amount necessary to improvesigns or symptoms a disease, such as osteoarthritis, a skin disorder, ora wound.

The NAG-containing food products and beverages disclosed herein haveequal application in medical and veterinary settings. Therefore, thegeneral term “subject being treated” is understood to include allanimals (such as humans, apes, dogs, cats, horses, and cows) thatrequire treatment of a cartilage dysfunction or skin disorder, such as awound.

Thermal tolerance: Refers to the ability of NAG to be exposed to a hightemperature, without a resulting significant adverse effect on thetaste, color, odor, or texture of a food or beverage supplemented withNAG, when NAG is present in the food or beverage during exposure to ahigh temperature. In particular examples, the amount of NAG present in aNAG food product or NAG beverage following exposure to a hightemperature, demonstrating that NAG is thermally tolerant, is at least70% of the original amount of NAG present, for example at least about75%, at least about 77%, at least about 80%, at least about 90%, atleast about 95%, at least about 98%, at least about 99%, or even 100%(no loss of NAG).

In contrast, food and beverage products including GLCN exposed to hightemperatures oftentimes have undesirable characteristics, such as anunpleasant taste and undesirable browning, when GLCN is present in thefood or beverage during exposure to a high temperature.

Treat: To alleviate or reduce one or more of the symptoms of a disorder,such as a cartilage dysfunction, wound or skin disorder, or to stabilizesuch a condition.

Advantages of NAG Over Other Cartilage Supplements

Certain embodiments of the disclosed food products and beveragesincluding NAG as a dietary cartilage supplement offer one or moreadvantages over the use of GLCN. In one example, the disclosed foodproducts and beverages containing NAG can be exposed to hightemperatures, such as those used in heat pasteurization or baking,without a significant adverse affect on the resulting taste, color,odor, or texture of the food or beverage. In certain examples, NAG foodproducts and NAG beverages are more stable at neutral or high pH valuesthan are similar products that include GLCN.

The nitrogen in NAG is in the neutral amide form, and therefore in someexamples has no substantial effect on the pH of the supplemented foodproduct or beverage. This pH tolerance makes NAG suitable for a widerange of foods and beverages, such as dairy products which can curdle atlower pH. In addition, because NAG is not a salt, in certain embodimentssubjects needing to lower their salt intake would not be adverselyaffected by consuming NAG-supplemented beverages or food products.Because NAG can impart sweetness to a supplemented food or beverage, insome examples it is used as a replacement for some or the entiresweetener in the food or beverage. For example, NAG can replace at leastabout 1% of a sweetener in a NAG food or NAG beverage, such as at leastabout 5% or at least about 10%. In certain embodiments, NAG does notparticipate in Maillard chemistry, and so does not contributeundesirable color to the product. Maillard chemistry is the nonenzymaticbrowning observed when amine-containing species, such as amino acids orproteins, react with carbohydrates during heating. Maillard chemistrycontributes to the browning obtained in many baked goods, such as breadand cookies.

Food and Beverages Supplemented with NAG

Disclosed herein are beverages and food products supplemented with NAG,and that are exposed to high temperatures, such as during processing orpreparation of the food or beverage. For example, NAG-supplemented foodproducts and beverages can be heat pasteurized or baked, withoutsignificant loss of NAG. For example, following exposure ofNAG-supplemented food products and beverages to high temperatures, theamount of NAG remaining in the food product or beverage is at least 70%,such as at least 90%.

The amount of NAG added to the beverage or food product will depend onthe desired concentration. In certain examples, NAG is present in thedisclosed food products and beverages in amounts effective for promotingthe development of connective tissue in the body, alone or incombination with other agents, such as cartilage promoting agents. Insome embodiments, daily NAG dosages include at least about 250 mg, atleast about 500 mg, at least about 1000 mg, at least about 2000 mg, oreven about at least 3000 mg. Particular NAG dosage ranges include, butare not limited to, a range of about 500 mg to about 3000 mg, such asabout 1000 mg to about 2000 mg, such as about 1500 mg of NAG.

Certain embodiments of the disclosed amounts of NAG that can be includedin a food or beverage include, at least about 0.001 g NAG/serving, suchas at least 0.01 g NAG/serving, at least about 0.05 g NAG/serving, atleast about 0.1 g NAG/serving, at least about 0.25 g NAG/serving, atleast about 0.5 g NAG/serving, at least about 0.75 g NAG/serving, atleast about 1 g NAG/serving, at least about 1.5 g NAG/serving, at leastabout 3.0 g NAG/serving, at least about 5 g NAG/serving, at least about10 g NAG/serving, or at least about 20 g NAG/serving. In other examples,the amount of NAG added is about 1 g NAG/1000 g of product to about 1 gNAG/0.1 g of product, such as about 1 g NAG/10 g product to about 1 gNAG/0.5 g product.

Certain embodiments of the disclosed NAG-supplemented food products andbeverages also include one or more other cartilage supplements,vitamins, minerals, fats, proteins, carbohydrates, sweeteners, organicacids, glucose or combinations thereof. In addition, other agents thattreat cartilage dysfunction or skin disorders can also be included inthe disclosed NAG-supplemented food products and beverages.

In some examples, the NAG food products and NAG beverages do not containdetectable amounts of shellfish proteins, such as those muscle proteinswhich are allergenic in some humans (that is, those persons havingshellfish allergies). In some examples, in order to decrease or eveneliminate the presence of shellfish proteins, NAG is derived from fungalbiomass, bacteria, or cartilage, instead of from shellfish.

In certain embodiments, NAG-supplemented beverages (or food products)can be heat pasteurized at a high temperature, wherein NAG is present inthe beverage (or food product) during the pasteurization. Particularnon-limiting heat pasteurization temperatures include, at least about160° F., at least about 180° F., at least about 200° F., at least about250° F., or at least about 300° F. Heat pasteurization in particularembodiments can also include exposure to high pressure, such as about121° C. at 1 atm for 15 minutes. In one example, NAG is included in acoffee, tea, or cocoa mixture (such as a pre-prepared packet) to whichboiling or heated water (or other liquid such as milk) is added.

NAG beverages, such as those including at least about 0.01 g NAG perserving, at a temperature of at least about 160° F., such as at leastabout 161° F., such as at least about 165° F., at least about 194° F.,at least about 200° F., at least about 212° F., at least about 220° F.,or even at least about 280° F., are encompassed by this disclosure.

Non-limiting examples of beverages that can be supplemented with NAGinclude naturally or artificially flavored fruit or vegetable juicessuch as apple juice, carrot juice, cherry juice, cranberry juice, grapejuice, grapefruit juice, orange juice, pear juice, tomato juice, or acombination thereof; milk; commercially available sports drinks (sugaror juice based) such as Gatorade®, Powerade®, and Allsport®; soda;Tang®; flavored waters; soy milk; and commercially availablenutritionally-balanced beverages such as Ensure® beverage. The beveragecan be carbonated or non-carbonated. In particular embodiments, thebeverage is in a concentrated form for later dilution by the consumer orready-to-drink. Alcoholic beverages are also encompassed by thisdisclosure, such as wine, wine coolers, malt beverages and coolers, andbeer.

Certain embodiments of the disclosed food products are thermallyprocessed after NAG is included in the food composition. Examples ofthermal processing include, but are not limited to, baking, roasting,broiling, and frying. Non-limiting examples of food products includeflour- and grain-based products, such as bakery products, for examplebread, cookies, muffins, rolls, brownies, pies and cakes (or mixes toprepare such products, such as cake mixes). Other non-limiting examplesinclude breakfast cereals, nutrition bars, snack bars, granola bars,animal feed products, bread crumbs, yogurt, gum, candy, andcanned-goods, such as soups, pastas, vegetables, fruits (such as piefillings) and meats.

In particular examples, NAG food products are exposed to a hightemperature, such as a temperature used in baking or frying, such as atleast about 300° F., at least about 325° F., at least about 350° F., atleast about 375° F., at least about 400° F., at least about 425° F., atleast about 450° F., and even such as at least about 500° F.

NAG food products, such as those including at least about 0.01 g NAG perserving and no detectable shellfish proteins, at a temperature of atleast about 160° F., such as at least about 161° F., such as at leastabout 165° F., at least about 194° F., at least about 200° F., at leastabout 212° F., at least about 300° F., at least about 350° F., at leastabout 400° F., or even at least about 500° F., are encompassed by thisdisclosure.

Methods of Preparing Foods and Beverages Supplemented with NAG

Methods of preparing beverages that include NAG are disclosed. In oneexample, the method includes heat pasteurizing the beverage, wherein NAGis present in the beverage during pasteurization. In another example,the method includes combining at least about 0.01 g NAG per serving anda beverage, thereby forming a NAG-beverage, and then exposing theNAG-beverage to heat pasteurization at a temperature of at least 160° F.

Also disclosed are methods for preparing NAG-supplemented food products.In one example, the method includes combining a food product and atleast about 0.01 g NAG per serving, wherein NAG is derived from fungalbiomass, bacteria, or cartilage such that no detectable shellfishallergens are present, and then exposing the NAG food product to a hightemperature, such as at least 160° F. For example, the NAG food productcan be baked; broiled; boiled; sterilized; canned; roasted; fried; suchas by heating in an oven, autoclave, microwave oven (or their industrialequivalents).

Treatment Using NAG

A method of treating a cartilage dysfunction in a subject byadministering the disclosed NAG-supplemented food products or beverages,is disclosed. In some examples, treatment alleviates or reduces thesymptoms of cartilage dysfunction, such as increases joint mobility,reduces pain or reduces swelling in the subject. In some examples,treatment stabilizes the symptoms of cartilage dysfunction, such thatthe cartilage dysfunction is not exacerbated. Examples of cartilagedysfunction include, but are not limited to, joint pain andosteoarthritis.

Also disclosed are methods of treating a sldn disorder in a subject byadministering the disclosed NAG-supplemented food products or beveragesto a subject. In some examples, treatment alleviates or reduces thesymptoms of a skin disorder, such as promotes wound healing in thesubject. In some examples, treatment stabilizes the symptoms of a skindisorder, such that the skin disorder is not exacerbated. Examples ofskin disorders include, but are not limited to, wounds and wrinkles.

A method for treating food allergies in a subject by administering thedisclosed NAG-supplemented food products or beverages to the subject isdisclosed. In some examples, treatment alleviates or reduces thesymptoms of a food allergy, such as reduces the inflammatory response tothe food in the subject. In some examples, treatment stabilizes thesymptoms of a food allergy, such that the food allergy is notexacerbated.

The subject treated can be a human or veterinary subject suffering fromcartilage dysfunction, skin disorder or food allergy (for example see WO93/14766A1). An effective amount of NAG can be administered in a singleserving, or in several servings, for example daily, during a course oftreatment. However, the effective amount can depend on the subject beingtreated, the severity and type of the condition being treated, and themanner of administration (food product versus beverage). A typicalamount of GLCN or NAG delivered in dietary supplement products is about1.5 g/day, in a single or in multiple administrations. For example, ifthe subject was to receive multiple administrations in a single day, thesubject might receive three servings of NAG, each containing about 0.5 gNAG. In certain embodiments, NAG is administered at about at least about0.01 g NAG/day, about at least 0.05 g NAG/day, about at least 0.1 gNAG/day, about at least 0.25 g NAG/day, about at least 0.5 g NAG/day,about at least 0.75 g NAG/day, about at least 1.0 g NAG/day, about atleast 1.5 g NAG/day, about at least 3.0 g NAG/day, about at least 5.0 gNAG/day, about at least 10.0 g NAG/day, or even about at least 20.0 gNAG/day.

EXAMPLE 1 Rice Krispies® Treats

Rice Krispies® Treats were used as the basis for incorporation ofsamples. According to the manufacturer, two treats are a serving. Toprepare the samples, four batches of Rice Krispies® Treats were preparedfor sensory testing as follows:

-   -   batch 1. Control (no GLCN, no NAG).    -   batch 2. GLCN (glucosamine hydrochloride derived from fungal        biomass, Cargill, Minneapolis, Minn., Lot No. GP015-A).    -   batch 3. GLCN (glucosamine hydrochloride derived from shellfish,        Batch 011107, Index # 9152, V.L. Clark Chemical Co., Inc.,        Union, Mo.).    -   batch 4. NAG (N-Acetyl-D-glucosamine, Lot # 27294A, Catalog #        A-102-2, Ferro Pfanstiehl, Waukegan, Ill.).

According to the manufacturer's instructions, the recipe prepares 24treats, so 4 batches were prepared from the recipe. Each batch consistedof six treats, or three servings. For servings which included GLCN orNAG, the serving (2 treats) had 0.75 g GLCN or NAG, or 0.375 g per treatadded, which reflects a typical amount of GLCN or NAG delivered indietary supplement products (0.25 g-1.5 g/serving). Therefore, exceptfor the control batch, each batch had 2.25 g of GLCN or NAG added (0.375g GLCN or NAG per treat×6 treats in batch=2.25 g GLCN or NAG per batch).GLCN or NAG was added to the melted marshmallow (melt marshmallow in a1000 watt Amana Radarange microwave following manufacturer'sinstructions) prior to adding the Rice Krispies®, to evenly disperse theminor dry ingredients. Here, the samples were not heated to hightemperatures.

Samples were tested within one day of preparing. Samples were testedblindly against a marked control, and a blind control was included.Panelists were asked to compare each sample to the control and comment.Panelists received the following instructions:

-   -   1. Taste the control sample first.    -   2. Compare all other samples to the control and write        descriptors in the table (some potential comments might be        whether or not the sample is hard, soft, brittle, crumbly,        chewy, or grainy).

3. Numerically rate whether the samples are better or worse than thecontrol, using this scale: −5 −4 −3 −2 −1 0 +1 +2 +3 +4 +5 Worse thancontrol Same as control Better than control

-   -   4. Please do not discuss the results with any other panelists        until all sheets are turned in.

The 12 Panelists' results are shown in Table 1. TABLE 1 Results of RiceKrispies ® Treats.* Mouthfeel/ Sample Color Odor texture Taste Comments(other than “same”) Control 0 −0.3 −0.4 0 A little crunchier (3) NAG 0+0.3 +0.3 +0.3 Much soggier, not good, shinier, chewier, sweeter (2)Fungal 0 −0.3 −0.1 −0.4 A little crunchier, softer (2), not good, slightsalt GLCN aftertaste, sour (3), slight aftertaste, sweeter, chewier,aftertaste, notice strange taste Shellfish 0 0 0 −1.4 Sour (3), softer(2), not good, chewier (3), tangy, GLCN slight aftertaste, sweeter,similar to aftertaste of cinnamon (burning), different but notunpleasant, aftertaste, stronger flavor*Results shown are the average rounded to nearest tenth.

As shown in Table 1, the results were validated by identification of thecontrol, based on the control score of near “0” in all categories. Inaddition, Table 1 demonstrates that NAG was preferred by the panelistsin Rice Krispies® Treats over either GLCN sample, and it wasunexpectedly favored over the control. These results demonstrate thatNAG has an advantage over GLCN when used in a high temperature foodapplication, as NAG does not appear to adversely affect taste afterbeing exposed to heated marshmallow.

EXAMPLE 2 Betty Crocker® Sugar Cookie Mix

A packet of Betty Crocker® dry cookie mix was used as the basis forincorporation of samples. A sugar cookie mix was used due to its lightcolor and mild taste. According to the manufacturer, two cookies are oneserving.

On package of sugar cookie mix prepares 36 cookies, so each batchconsisted of nine cookies, or 4.5 servings. For servings which includedGLCN or NAG, the serving (2 cookies) had 0.75 g GLCN or NAG, or 0.375 gper cookie added, which reflects a typical amount of GLCN or NAGdelivered in dietary supplement products. Except for the control, toeach batch 3.375 g of NAG or GLCN was added (0.375 g GLCN orNAG/cookie×9 cookies in batch=3.375 g GLCN or NAG/batch). Each batch wasbaked separately, so that the cookies were baked in the same places onthe cookie sheet. GLCN or NAG was added to the dry mix and mixed with aspoon. Before baking each batch, 1/12 cup of oil, followed by ¼ of abeaten egg, was added. The mixture was stirred by spoon, then by hand,to form a soft dough. The dough was weighed and divided into nine evenballs and placed at least two inches apart on an ungreased cookie sheet,then flattened in a crisscross pattern with a fork. Cookies were bakedat 375° F. for 8-10 minutes on a shiny metal pan. Cookies were cooledfor two minutes before removing from cookie sheet, then stored inairtight containers.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 2. TABLE 2Results of Betty Crocker ® Sugar Cookies.* Mouthfeel/ Sample Color Odortexture Taste Comments (other than “same”) Control −0.3 −0.2 0 −0.1Darker around edges, harder NAG 0 +0.1 +0.2 −0.1 Slightly sweeter, moremoist, chewier (3), looks lighter than control, crumbly, smells sweeterFungal −1.5 −0.3 −0.7 −1.7 Crumbly (2), burnt sugar/caramel taste (4),GLCN darker and/or dark specs on bottom (10), slight cinnamon-like hottaste, strange aftertaste, taste & smell sweet, crunchier/harder (4),toasted odor, off smell, grey/brown border on bottom is unappealing (2),off/bitter aftertaste-almost salty Shellfish −1.0 −0.1 −0.2 −0.8 Crumbly(2), burnt taste, chewier, dark specs GLCN throughout and on bottom (8),smell sweet, taste sweet (2), chewy, crunchier, toasted odor, flourtaste, color change on border is unappealing, firmer*Results are the average rounded to nearest tenth.

The results shown in Table 2 were validated by identification of thecontrol, based on the control score of near “0” in all categories. Asshown in Table 2, NAG was preferred by the panelists in sugar cookiesover either GLCN sample, and the scores indicate it was even favoredover the control. Only the GLCN samples showed significant darkening,indicating adverse interactions between the food components and GLCNduring heating. Therefore, panelists did not like the taste orappearance of the GLCN samples, and the NAG sample was accepted orfavored. These results demonstrate that NAG has an advantage over GLCNwhen used in a high temperature food application, as NAG does not appearto adversely affect taste or browning after being heated to 350° F.,unlike GLCN. Because NAG enhanced the sweetness of the sugar cookies,NAG can be used to replace all or some of the sweetener, such as atleast 5%, at least 10%, or at least 20% of a sweetener used in abeverage or food product.

EXAMPLE 3 Allsport® Sports Drink

Allsport® sports drink (The Monarch Company, Atlanta, Ga.) was used asthe basis for incorporation of samples. Citrus Slam flavor was chosenbecause of its light color, which may help panelists observe any colorchange. According to the manufacturer, a 20-ounce bottle of Allsport®contains 2.5 servings.

For servings that included GLCN or NAG, the serving had 0.75 g GLCN orNAG added, which reflects a typical amount of GLCN or NAG delivered indietary supplement products. Therefore, 1.875 g (0.75 g/serving GLCN orNAG×2.5 servings/bottle=1.875 g GLCN or NAG/bottle) of GLCN or NAG wasadded to a 20-ounce bottle of Allsport®. After the bottles wereprepared, the contents were heated to 195±4° F. for 42±4 seconds tosimulate a pasteurization step. Samples were heated in a 1000 watt AmanaRadarange microwave oven in foam cups for the same amount of time. For aroom temperature beverage, 150 seconds of heating time was necessary,and for a refrigerated beverage, 180 seconds was required. The beverageswere cooled in their original bottles, ¾ submerged in an ice water bath,and hand rotated to simulate flash cooling.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 3. TABLE 3Results of Allsport ® sports drink.* Mouthfeel/ Sample Color Odortexture Taste Comments (other than “same”) Control +0.1 0 +0.4 −0.1 Lesssour than others, sweeter (2), more of a carbonated feel, stronger odor(2), more tangerine taste, mouthfeel thicker NAG −0.1 +0.1 0 −0.2 Lesstangy/sour than others (3), sweeter, liked it better, crystals on topFungal 0 −0.3 +0.1 −0.3 After/offtaste (2), “vitamin” like taste, GLCNsweeter, less odor, sour, yucky Shellfish −0.1 −0.3 −0.2 −0.7 Sweeter(2), sour, crystals on top, less odor, GLCN mouthfeel thicker, lesstangy, flat and yucky*Results are the average rounded to nearest tenth.

As shown in Table 3, it was difficult for the panelists to ascertaindifferences between the samples. The control was not clearlydifferentiated from the NAG or fungal GLCN. However, the NAG sampleswere favored over both GLCN samples. These findings further demonstratethat taste and other factors are not adversely affected when NAG is usedin a high temperature beverage application, such as those that requirepasteurization. Further distinctions between NAG and GLCN may beobserved if higher temperatures or extended times are used.

EXAMPLE 4 Nestle Toll House® Milk Chocolate Morsels

Nestle Toll House® Milk Chocolate Morsels were used as the basis forincorporation of samples. One serving of morsels is 14 g.

The shellfish GLCN was more granular than the other samples, so it wasmilled to disperse better in the chocolate. For servings which includedGLCN or NAG, the serving (14 g) had 0.75 g GLCN or NAG added, whichreflects a typical amount of GLCN or NAG delivered in dietary supplementproducts. As 217 g of morsels were used for the batches which had GLCNor NAG, 11.625 g of GLCN or NAG was added (217 g is 15.5 servings, 15.5servings×0.75 g/serving=11.625 g). Each batch was heated in a 1000 wattAmana Radarange at medium-high power for one minute. The sample wasstirred and again heated in the microwave for additional 20 secondintervals until the chocolate was smooth. The temperature of thechocolate was recorded with a candy thermometer, and the GLCN or NAGadded to the appropriate batches and stirred in quickly and thoroughly.Plastic spoons were used to drop teaspoon-sized amounts of chocolateonto wax paper, and were allowed to cool.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 4. TABLE 4Nestle Toll House ® Milk Chocolate Morsels results.* Mouthfeel/ Comments(other Sample Color Odor texture Taste than “same”) Control 0 0 0 +0.2Dry, rough, creamy NAG −0.8 −0.3 −1.4 −0.9 Dry/crumbly (4), roughsurface (4), sticky in mouth, crunchy/grainy (2), flavor better thancontrol (3), “thicker” taste not good, less sweet, off flavor, bitterFungal 0 −0.1 −0.8 −2.0 Bitter (7), sour (2), GLCN grainy (4), salty(2), burnt taste, metallic, strange aftertaste Shellfish 0 −0.1 −2.8−2.5 Crunchy/gritty (8), GLCN sour (2), bitter (4), salty (2), badaftertaste*Results are the average rounded to nearest tenth.

As shown in Table 4, the results were validated; based on the controlscore of near “0” in all categories, the control was identified. Thetaste of NAG was preferred by the panelists in milk chocolate overeither GLCN sample, and three panelists favored it over the control.Both GLCN samples were bitter or sour and grainy. The appearance of theNAG sample was less desirable, likely because when the NAG was added tothe chocolate, it thickened the texture, making it more like fudge.These results demonstrate that NAG has an advantage over GLCN when usedin a high temperature food application, as NAG does not appear toadversely affect taste after being heated, unlike GLCN.

EXAMPLE 5 Libby's White Grape Juicy Juice®

Libby's White Grape Juicy Juice® was used as the basis for incorporationof samples. White Grape was chosen due to its light color, to helppanelists observe any color change. According to the manufacturer, aserving is 8 ounces.

For servings that included GLCN or NAG, the 8 ounce serving had 0.75 gGLCN or NAG added, which reflects a typical amount of GLCN or NAGdelivered in dietary supplement products. A 64 ounce bottle of juice wassplit into four 16-ounce portions. To the appropriate batch, 1.5 g ofGLCN or NAG (16 ounces is two servings, and 2 servings×0.75g/serving=1.5 g) was added. After preparing the samples, they wereheated to 195±4° F. to simulate a pasteurization step in a microwave,then cooled, as described in Example 3.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 5. TABLE 5Libby's White Grape Juicy Juice ® results.* Mouthfeel/ Comments (otherSample Color Odor texture Taste than “same”) Control +0.1 0 0 0 Slightoff taste, more lingering taste NAG −0.3 0 −0.1 −0.5 Sweeter, less sweet(2), rouge color, bitter (2), darker (3), lingering taste Fungal −0.2−0.3 −0.3 −0.6 Darker (2), thicker GLCN mouthfeel (2), sweeter (2),stale/weaker (2), smooth, soapy, lasting aftertaste, tarty, odor maskedShellfish −0.4 −0.1 −0.3 −1.1 Darker (3), thicker GLCN mouthfeel,sweeter (2), soapy (2), tarty, bitter, less sweet, less fruity (2)*Results are the average rounded to nearest tenth.

The results shown in Table 5 were validated by identification of thecontrol, based on the control score of near “0” in all categories. Asshown in Table 5, it was difficult for the panelists to ascertaindifferences between the samples. However, NAG was preferred over theGLCN samples, but not overwhelmingly so. These results demonstrate thattaste and other factors are not adversely affected when NAG is used in ahigh temperature beverage application, such as those including apasteurization step.

EXAMPLE 6 Caramels

Made from scratch caramels were used as the basis for incorporation ofsamples. One serving is two caramels. For servings which included GLCNor NAG, the serving (2 caramels) had 0.75 g GLCN or NAG, or 0.375 g percaramel added which reflects a typical amount of GLCN or NAG deliveredin dietary supplement products. Batches of 8 servings were prepared. Forservings that included GLCN or NAG, 6 g GLCN or NAG was added (8servings×0.75 g/serving=6 g).

A Better Homes & Gardens recipe was used. To make the caramels, ¼ cupnon-corn oil margarine was melted over low heat in a nonstick-coatedsaucepan. Packed brown sugar (½ cup), ½ cup half and half (substitutedfor light cream), and ¼ cup light corn syrup was added and mixed well.GLCN or NAG was added to the appropriate batches, and the samplesstirred well. The samples were cooked and stirred over medium-high heatto boiling. The mixture was cooked and stirred over medium heat to 248°F. until a firm-ball stage was reached. The saucepan was removed fromthe heat, ¼ teaspoon imitation vanilla added to the mixture, then themixture immediately poured into a pan lined with buttered-foil and allowto cool.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 6. TABLE 6Caramel results.* Mouthfeel/ Sample Color Odor texture Taste Comments(other than “same”) Control 0 0 0 +0.1 NAG 0 −0.2 −0.3 −0.2 Lightercolor, smell is masked, more grainy (3), sweeter, a little better thancontrol, didn't taste as buttery, a little aftertaste, not as sweet (3),not as chewy Fungal −1.7 −0.3 −0.5 −3.5 Darker in color (7), burntcaramel taste (2), harder GLCN than control (2), salty and solventtaste, tasted bad, bitter (7), not very pleasant, reduced caramel andbutter flavor, tart, too tangy, less sweet odor, burnt odor, sourShellfish −1.4 −0.3 −0.9 −3.5 Darker in color (7), burnt caramel taste,salty and GLCN solvent taste, tasted bad, sour (2), after taste hitshard, acidic tart, suppressed caramel & buttery flavor, almost rancid orsalty, less sweet odor, bitter (7), burnt odor*Results are the average rounded to nearest tenth.

The results shown in Table 6 were validated; based on the control scoreof near “0” in all categories, the control was identified. As shown inTable 6, the taste of NAG was preferred to either GLCN sample, and NAGrated at or near control levels. Although three panelists did note thatthe NAG sample was more grainy and less sweet, the scores do not reflectthat was perceived as highly unfavorable. These results demonstrate thatNAG has an advantage over GLCN when used in a high temperature foodapplication, as NAG does not adversely affect taste or browning afterbeing heated as does GLCN.

EXAMPLE 7 Jiffy® Corn Muffin Mix

Jiffy® Corn Muffin Mix was used as the basis for incorporation ofsamples. Corn muffins were chosen due to their light color and mildtaste. One serving is one muffin (as per the manufacturer).

Two boxes prepare 12 large muffins, a box was used to prepare 4 batchesof samples (3 muffins per batch). For servings which included GLCN orNAG, the serving (1 muffin) had 0.75 g GLCN or NAG added, which reflectsa typical amount of GLCN or NAG delivered in dietary supplementproducts. Except for the control, each batch had 2.25 g of NAG or GLCNadded (0.75 g NAG or GLCN/muffin×3 muffins=2.25 g NAG or GLCN/batch).Each batch was prepared separately, so that the muffins were baked inthe same places on the muffin pan. GLCN or NAG was added to the dry mixand mixed with a spoon. To this, 4 teaspoons of milk and ¼ of a beatenegg were added and the ingredients blended until slightly lumpy (eachbatch was mixed for the same amount of time). The mixtures were allowedto sit for three minutes, then evenly distributed into lightly-greasedmuffin cups. The muffins were baked at 400° F. for 18 minutes, thenimmediately removed from the muffin cup and stored in airtightcontainers.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 7. TABLE 7Jiffy ® Corn muffin results.* Mouthfeel/ Sample Color Odor texture TasteComments (other than “same”) Control −0.3 −0.1 0 −0.3 Darker (2), smellssweeter, tastes sweeter, bitter, drier NAG +0.3 −0.2 −0.1 −0.1 Not goodtexture, slight aftertaste, lighter color (2), more sweet/less corntaste, more flour texture, less odor, better than control Fungal −2.1−0.3 −0.8 −2.4 Darker (8), dark specs (2) crunchy, grainy, GLCN drier,bitter (4), unpleasant taste (6), burnt smell, soapy mouthfeel, lessodor (3), chewier, salty/solvent taste, sour milk taste, sweeterShellfish −1.7 −0.4 −0.1 −1.0 Darker (6), dark specs (5), alum liketaste, GLCN bitter (3), less odor (2), burnt odor, “fresher”, more flourtexture (2), less taste, less sweet, grainy, sour, drier*Results are the average rounded to nearest tenth.

The results shown in Table 7 were validated; based on the control scoreof near “0” in all categories, the control was identified. As shown inTable 7, the taste of NAG was preferred by the panelists in corn muffinsto either GLCN sample. NAG performed at or slightly better than thecontrol. Differences in taste were noticed by a few panelists in thecrusts vs. the non-crust portion of muffins. These results demonstratethat NAG has an advantage over GLCN when used in a high temperature foodapplication, as NAG does not appear to adversely affect taste orbrowning after being heated, unlike GLCN.

EXAMPLE 8 Hy-Vee® Fudge Brownie Mix

Hy-Vee® Fudge Brownie Mix was used as the basis for incorporation ofsamples. Brownies, which have a darker color, were used to mask anybrowning from panelists as browning could cause bias by the obviouspresence of a non-control sample. According to the manufacturer, one boxmakes 15 brownies (1 serving is 1 brownie). To prepare the samples, thedry mix was separated into four batches, each with 3.75 servings.

For servings which included GLCN or NAG, the serving (1 brownie) had0.75 g GLCN or NAG added, which reflects a typical amount of GLCN or NAGdelivered in dietary supplement products. Except for the control GLCN orNAG was added to each batch (0.75 g NAG or GLCN/brownie×3.75brownies=2.8 g NAG or GLCN/batch). Each batch was prepared separately,so that the brownies were baked in the same places on cookie sheet. The“Quick Brownie Cookie” recipe on the box was used. GLCN or NAG was addedto the dry mix and mixed with a spoon. To each batch, 4 teaspoons coolwater, 2¼ teaspoons vegetable oil and ¼ beaten egg was added. The mixwas stirred with a spoon (about 40 strokes) until smooth and shiny. Themixture was dropped by teaspoonfuls onto a greased cookie sheet andbaked for 10 minutes at 350° F. After baking, the brownies wereimmediately removed from the cookie sheet, allowed to cool, and storedin airtight containers.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 8. TABLE 8Hy-Vee ® Fudge Brownie results.* Mouthfeel/ Sample Color Odor textureTaste Comments (other than “same”) Control 0 0 −0.1 0 Slightly drier,odor masked, less chewy NAG −0.6 −0.4 −0.6 −0.7 Chewy (3), exteriorshiny (2), reddish color, odor masked (2), less sweet, more chocolatetaste, smoother texture, taste off a little but not too bad, darker,smells salty/burnt, tastes burnt Fungal −3.1 −2.3 −3.6 −4.1 Roughtexture, strong aftertaste (2), sour (2), GLCN bitter (3), tangy, burnttaste (5), burnt (8), burnt smell (3), sour odor, crispy (7), darker(6), dry, sticks to teeth Shellfish −0.4 −0.8 −1.3 −2.7 Strongaftertaste (2), smooth texture, chewy GLCN (4), sour (2), bitter (3),darker, reddish color, exterior shiny, bitter smell, rancid smell,rancid taste, crispy granules (2), less sweet, darker, masked odor,taste salty/burnt, burnt taste*Results are the average rounded to nearest tenth.

The results in Table 8 were validated; based on the control score ofnear “0” in all categories, the control was identified. As shown inTable 8, taste of NAG was preferred by the panelists more than eitherGLCN sample. These results demonstrate that NAG has an advantage overGLCN when used in a high temperature food application, as NAG does notappear to adversely affect taste or browning after being heated to 350°F., unlike GLCN.

EXAMPLE 9

Fleischmann's® Country White Bread Machine Mix

Fleischmann's® Bread Machine Mix was used as the basis for incorporationof samples. Country White Mix was chosen due to its light color and mildflavor. According to the manufacturer, 1 box make 8 servings (8 slices).

One box of bread mix was used for each batch. For servings whichincluded GLCN or NAG, the serving (1 slice) had 0.75 g GLCN or NAGadded, which reflects a typical amount of GLCN or NAG delivered indietary supplement products. Except for the control, 6 g GLCN or NAG wasadded to each batch (0.75 g NAG or GLCN/slice×8 slices=6 g NAG orGLCN/batch). The recipe on the box was used. GLCN or NAG was added tothe dry mix and mixed with a spoon, and the mixture added to abreadmaker. To this, 8 ounces of water (75° F.-85° F.), and 1 package ofyeast (provided with the bread mix) was added. The bread machine was setto medium/normal crust color. The finished bread was immediatelyremoved, cooled on a plate, and stored in airtight containers.

Samples were tested according to the methodology and instructions inExample 1. The 10 Panelists' results are shown in Table 9. TABLE 9Fleischmann's ® Country White Bread Machine Mix results.* Mouthfeel/Sample Color Odor texture Taste Comments (other than “same”) Control 0 0−0.3 −0.2 Less odor, drier (2) NAG −0.1 0 0 −0.3 Sour, darker, darkercrust, tougher, heavier texture, more porous (2), salty/briny aftertasteFungal −0.9 −0.3 −1.1 −1.0 Doughy (4), darker (2), darker crust, darkerbread, GLCN denser (3), spongier, chewier (4), earthy taste, sour,sweeter (2), bitter, moister (3), “breadier” smell, crust slightlycrisp, yeasty, gummy after chewed Shellfish −0.9 −0.2 −0.4 −1.1 Darker(2), darker crust, darker bread, sour, GLCN spicy/musty odor, denser,spongier (2), earthy taste, bitter taste (2), burnt taste, vinegartaste, moister, “breadier” smell, chewier, more bread taste*Results are the average rounded to nearest tenth.

The results shown in Table 9 were validated; based on the control scoreof near “0” in all categories, the control was identified. As shown inTable 9, the taste of NAG was preferred by the panelists more thaneither GLCN sample, with NAG performing about as well as the control.These results demonstrate that NAG has an advantage over GLCN when usedin a high temperature food application, as NAG does not appear toadversely affect taste or browning after being baked at hightemperatures, unlike GLCN.

EXAMPLE 10 Hy-Vee® Healthy Recipe Tomato Soup

Hy-Vee® Healthy Recipe Tomato Soup was used as the basis forincorporation of samples. Tomato soup was chosen for its even texture,and “healthy” aspect as compared to other soups. According to themanufacturer, one can makes 2.5 servings.

For servings that included GLCN or NAG, the serving had 0.75 g GLCN orNAG added, which reflects a typical amount of GLCN or NAG delivered indietary supplement products. Each batch prepared 2.5 servings, so exceptfor the control, GLCN or NAG was added to each batch (0.75 g GLCN orNAG/serving×2.5 servings=1.875 g GLCN or NAG/batch). The recipe on thecan was used. The soup can contents were poured into a microwaveablecontainer, one can of water slowly stirred in, and GLCN or NAG added(where applicable) and the mixture stirred well. The container wascovered with vented plastic wrap (turn back edge of wrap to form smallopening for steam to escape), and the mixture heated in a 1000 wattAmana Radarange at high power for 2 minutes. The sample was not allowedto come to a boil.

Samples were tested according to the methodology and instructions inExample 1. The 8 Panelists' results are shown in Table 10. TABLE 10Hy-Vee ® Healthy Recipe Tomato Soup results.* Mouthfeel/ Comments SampleColor Odor texture Taste (other than “same”) Control −0.2 0 −0.1 +0.2less tangy (2), lighter in color NAG 0 0 0 0 Sweeter, less acidic withweaker aftertaste Fungal 0 0 −0.1 −0.3 Off flavor, a little GLCNmetallic, flat/sour taste but barely perceptible, more tangy w/acidicaftertaste (like black pepper had been added), milder, maybe sweeter,less tangy aftertaste Shellfish −0.2 0 −0.3 −0.6 Slight off flavor,dirty GLCN odor, flat/sour taste but barely perceptible, lighter incolor, off aftertaste, bitter, chalky in texture, sour*Results are the average rounded to nearest tenth.

The results shown in Table 10 were validated by identification of thecontrol; based on the control score of near “0” in all categories, withNAG scoring as well as the control. Even though it was difficult for thepanelists to ascertain differences between the samples, the NAG wasfavored over the GLCN samples. These results further demonstrate thattaste and other factors are not adversely affected when NAG is used in ahigh temperature food application, such as those which include heatingprior to consumption.

EXAMPLE 11 Bubble Gum

Canny Kits® Bubble Gum was be used as the basis for incorporation ofsamples. A serving of gum was estimated to be two pieces of gum.

According to the manufacturer's instructions, the mix makes over ¼ poundof gum. Assuming a piece of gum weighs at least 4 g, the mix can prepareapproximately 30 pieces of gum. A gum kit was used to make two batchesof gum. For servings which included GLCN or NAG, the serving (2 piecesof gum) had 0.75 g GLCN or NAG, or 0.375 g GLCN or NAG per piece of gumadded, which reflects a typical amount of GLCN or NAG delivered indietary supplement products. Except for the control batch, 5.625 g ofGLCN or NAG was added (0.375 g active per piece of gum×15 pieces of gumin ½ a gum kit=5.625 g).

To make the gum, a plate was covered with waxed paper. Half of thepowdered sugar provided was poured onto the waxed paper, and a well wasformed in the pile of sugar to receive the melted gum and syrup. Thegum-base pellets were melted by placing them in a plastic cup with thecorn syrup, flavoring and GLCN or NAG (as applicable), then microwavingin a 1000 watt Amana Radarange microwave on high for 15 seconds. Theheating step was repeated for two more 15 second intervals, stirringafter each, until the gum base was completely melted. This meltedmixture was slowly added to the powdered sugar, with constant mixing.The gum was worked into the powdered sugar until it no more sugar wouldabsorb. When the gum was cool enough to handle, it was handled likebread dough and rolled flat, cooled, and cut into pieces for sensorytesting.

Samples were tested according to the methodology and instructions inExample 1. The twelve Panelists' results are shown in Table 11. TABLE 11Results of Canny Kits ® Bubble Gum.* Mouthfeel/ Comments Sample ColorOdor texture Taste (other than “same”) Control +0.1 −0.1 Not as sweet,good NAG +0.1 −0.2 −0.3 Drier (2), crumblier, weaker texture, lessfruity taste, waxy, not as soft, metallic aftertaste Fungal −0.3 −0.6−2.9 Grainy, bitter (4), salty GLCN (3), bad, darker stale musky tasteShellfish −0.2 −0.8 −1.8 Grainy (2), bitter (2), GLCN stickier, salty(2), good, harder, stale musky taste, less fruity taste*Results are the average rounded to nearest tenth.

As shown in Table 11, the results were validated; based on the controlscore of near “0” in all categories, the control was identified. Thetaste of NAG was preferred by the panelists in gum over either GLCN, andthe NAG was rated at or near control levels. These results demonstratethat NAG has an advantage over GLCN when used in a high temperature foodapplication, as NAG does not appear to adversely affect taste afterbeing heated, unlike GLCN.

EXAMPLE 12 Tea

Tea made from Lipton® instant tea mix was used as the basis forincorporation of samples. To prepare the tea, the following recipe wasused: 2.69 g instant tea, 5.7 g GLCN or NAG (where applicable) then addwater to one quart. Tea was prepared for sensory testing as follows:

Batch 1. Control Tea (no NAG or glucosamine)

Batch 2. Tea+glucosamine (pH 5-6) [GLCN from fungal biomass, see batch 2Example 1]

Batch 3. Tea+NAG (pH 5-6) (N-Acetyl-D-glucosamine, Catalog # A-102-2,Ferro Pfanstiehl, Waukegan, Ill.)

Batch 4. Tea+glucosamine (pH 5-6) heated to 71° C. for 20 minutes

Batch 5. Tea+glucosamine heated (pH 5-6) to 90° C. for 20 seconds, flashcooled in ice water bath

Batch 6. Tea+glucosamine (pH 5-6) boiled on stove for 5 minutes, flashcooled in ice water bath

Batch 7. Tea+NAG (pH 5-6) heated to 71° C. for 20 minutes

Batch 8. Tea+NAG (pH 5-6) heated to 90° C. for 20 seconds, flash cooledin ice water bath

Batch 9. Tea+NAG (pH 5-6) boiled on stove for 5 minutes, flash cooled inice water bath

The recipe prepared one quart, so each batch consisted of 32 ounces, orfour 8-ounce servings. For servings that included GLCN or NAG, theserving (8 ounces) had 1.425 g GLCN or NAG, or 5.7 g GLCN or NAG perbatch (1.425 g/serving×4 servings/batch=5.7 g/batch), which reflects atypical amount of GLCN or NAG delivered in dietary supplement products.

Samples were tested within one day of preparing. Samples were testedblindly against a marked control, and a blind control was included.Panelists were asked to compare each sample to the control and commentusing the instructions and scale described in Example 1. The sixpanelists' results are shown in Table 12. TABLE 12 Results of Tea.*Actual Initial pH after 30 sec Sensory Panel Sample Temp ° C. pH heatingTemp ° C. Mouthfeel Taste Observations Control Not 5.47 N/A N/A 0.7 3.0Normal, like heated Lipton, slightly dry GAP Not 5.09 N/A N/A −0.7 −3.2Salty, syrupy heated texture, sweeter, bitter GAP 71 5.09 5.13 69 −0.5−3.7 Sweet aftertaste, 71° C. bitter, salty Tea + GAP 90 5.09 5.00 78−0.5 −4.2 Salty (2), sweet, 90° C. sour (2), bitter, a little darker GAP94 5.09 4.87 74 −0.7 −2.2 Sweeter (2), boiled bitter, astringent, alittle darker NAG Not 5.46 N/A N/A 0.7 2.0 Like control, heated sweet(2), bitter Tea + NAG 71 5.46 5.40 67 −0.2 1.3 sweet 71° C. NAG 90 5.465.31 81 −0.2 1.2 Bitter, tangy 90° C. NAG 96 5.46 5.22 74 −0.3 0.2 Verybitter, boiled caramel like but not sweet, darker*Values shown are the average rounded to nearest tenth.

As shown in Table 12, NAG was preferred over GLCN in tea, which was moreneutral tasting than the lemonade. The pH of the tea shows that the pHslightly decreased after heating in most cases (with both GAP and NAG).

EXAMPLE 13 Comparison of GLCN-HCl and GLCN-Sulfate

As described in the above, examples, use of glucosamine-hydrochloride infood and beverages heated to high temperatures, often resulted in a souror bitter aftertaste. To determine if similar results would be obtainedwhen glucosamine-sulfate was used, the methods described in Example 2(sugar cookies) were repeated using a GLCN-sulfate sample in addition toa GLCN HCl sample.

GLCN-sulfate (D-Glucosamine Sulfate 2KCl) was obtained from AnhuiTechnology Import & Export Co., Ltd. (Hefei, P.R. China, Batch #2002KO925, manufacturing date: Sep. 24, 2002). Cookies were prepared asdescribed in Example 2.

Samples were tested according to the methodology and instructions inExample 1. The 12 Panelists' results are shown in Table 13. TABLE 13Effect of GLCN-Sulfate versus GLCN-HCl.* Mouthfeel/ Sample Color Odortexture Taste Comments (other than “same”) Control −0.1 −0.1 −0.2 −0.2NAG −0.2 −0.2 −0.8 −0.2 Darker(2), more vanilla odor, less odor, lesssweet, sweeter(3), crispier or crunchier(5), less buttery taste, saltytaste, chewy Fungal −2.5 −1.8 −1.1 −3.1 burnt(2), darker(7), doughy(2),less sweet odor, GLCN softer(2), burnt taste, bitter taste(2), sourtaste(2), black specs, salty taste, burnt sugar taste (2), molassestaste, less sweet taste, tangy taste, bitter odor, pungent dirty odor,chewy GLCN −1.6 −0.8 −1.0 −2.0 Darker(5), black specs(3), doughy, lessodor, sulfate softer(5), “melts” in mouth, cinnamon “hot” aftertaste,sour taste, salty taste, bitter taste, tangy taste, bitter odor, sweetertaste, chewy, less buttery odor, less buttery taste*Results are the average rounded to nearest tenth.

The results shown in Table 13 are validated by identification of thecontrol; based on the control score of near “0” in all categories. NAGwas liked better by the panelists in sugar cookies than either GLCNsample, and it scored almost the same as the control. Only the GLCNsamples showed significant darkening, indicating adverse interactionsbetween the food components and GLCN during heating. These resultsdemonstrate that NAG has an advantage over GLCN (HCl and sulfate) whenused in a high temperature food application, as NAG does not appear toadversely affect taste or browning after being baked at hightemperatures, unlike GLCN. In addition, it appears that both forms ofGLCN (HCl and sulfate), have undesirable properties when heated.

EXAMPLE 14 Simulation of Stomach Conditions

As described in the above examples, NAG does not impart off flavors andcolors to the extent observed by glucosamine, when used as a minor(dietary supplement) ingredient in high temperature applications. Todemonstrate that NAG is biologically available once in the stomach, thefollowing methods were used.

To test the availability of NAG in the stomach, a simulated stomachenvironment was constructed, and a known concentration of NAG was addedto simulated gastric fluid (SGF) and maintained at conditions simulatingthe stomach. If NAG is degraded or derivatized in the stomach, the NAGconcentration will decrease. If the concentration of NAG remains thesame, it is likely that the NAG is available to the body.

SGF was prepared based on a 1995 United States Pharmacopoeia (USP)monograph. An artificial stomach environment was simulated according togenerally accepted practice (see Consumers Union website, 101 TrumanAve., Yonkers, N.Y. 10703-1057). To prepare simulated gastric fluid, 2.0g of NaCl and 3.2 g of pepsin were dissolved in 7.0 mL of hydrochloricacid (concentrated) and sufficient water to make a 1000 mL solution.This solution had a pH of about 1.2.

NAG samples were prepared as follows. A typical amount of GLCN or NAGdelivered in dietary supplement products is 0.5 -1.5 g/serving, and isoften taken in multiple 0.5 g or 0.75 g doses, or as a single dose. NAG(1 g) was placed in the simulated stomach (SGF solution) to be at anoptimum concentration for the instrument, and to reflect a reasonableingestion of NAG as a supplement.

To simulate the stomach, experiments were conducted at 37° C., which isnormal human body temperature, in a water bath/shaker, to gently agitatethe samples. Beakers sealed with parafilm were used to restrict loss ofsample.

Before introduction of samples into the SGF, a sample of the SGF wasanalyzed as a blank, to determine the baseline. Upon addition of the NAGto the SGF, a sample was immediately analyzed to determine theconcentration of NAG at time zero. After introduction of NAG into theSGF, samples were pulled and analyzed at 5 minutes, 15 minutes, and 60minutes to provide a profile of NAG concentration in the simulatedstomach, and analyzed for the concentration of NAG.

After subjecting the NAG to the SFG as described above, there was nosignificant loss of NAG with respect to the accuracy of the method.Therefore, it is likely that the NAG is available to the body afteringestion, since it does not appear that NAG is significantly chemicallyaltered (such as degraded or derivatized) following ingestion.

EXAMPLE 15 Determination of the Amount of NAG Present following Heating

To demonstrate that NAG remains following heating or pasteurization offood products and beverages, the following methods were used. Foodsamples were homogenized prior to analysis. Where possible, samples werefrozen or dried and ground. Baked goods were crumbled and mixed toproduce a homogeneous material. Suitable blanks (samples with no addedNAG) of each food type were analyzed to assess interferences. Masschanges between unheated ingredient mixtures and baked final productswere tracked to permit accurate recovery calculations.

Acid Extraction Method

A food or beverage sample as described in the preceding examplescontaining 5 to 20 mg of N-acetylglucosamine (NAG) was dispersed in 25 gof 0.1 N HCl in a 50-mL polypropylene centrifuge tube and cappedtightly. The sample was mixed for 30 seconds using a vortex mixer, thenplaced in a water bath at 37° C. The sample was removed from the waterbath at 15-minute intervals and mixed for 30 seconds on a vortex mixerand then returned to the water bath. This cycle was repeated until thesample had been in the water bath for one hour. After heating, thesample was mixed for 30 seconds on a vortex mixer, then centrifuged for10 minutes to separate the liquid and solid phases. Fats, oils or lipidsin the sample formed a third layer at the top of the tube. The aqueousportion of the sample was filtered through a 0.2μ filter into an HPLCvial, then capped.

NAG recovery was determined using high performance liquid chromatography(HPLC) using a combination of refractive index and UV (195 nm)detection. The system included a SIL-10AXL autosampler, SCL-10AVPcontroller, LC-10AT pump, CTO-6A column oven, SPD-M10AVP diode-arraydetector, and a RID-6A refractive index detector, all from ShimadzuScientific Instruments, Inc. (Columbia, Md.). The column was a MetaCarbH Plus, 300×7.8 mm, from Varian, Inc. (Torrence, Calif.).

The eluent, 0.01N sulfuric acid in water, flow rate was 0.4 mL/min. Thecolumn was maintained at 70° C. A 10 μL injection volume was used. NAGeluted at 23.9 minutes and was well resolved from other species in thesamples. Multiple standards confirmed good linearity over theconcentration range of interest. The LW spectrum from 190 to 350 nmindicated no measurable co-eluting peaks, and the retention time andratio of responses between the detectors confirmed the identity of NAG.

As shown in Table 14, the amount of NAG recovered following heatingranged from about 77%-100%.

AOAC Method

One method used to determine the amount of NAG in processed food sampleswas adapted from “Glucose, Fructose, Sucrose, and Maltose inPresweetened Cereals: Liquid Chromatography Method”, AOAC Method 982.14,15^(th) Ed. (1990), pp. 789-790 (herein incorporated by reference).Specifically, section C of the method was adapted to extract NAG fromdry-mixed and baked samples.

The sample was dried (if needed) then ground to render it homogeneous.Approximately five grams of sample were mixed with 100 mL of a 1:1water:ethanol solution. The samples were heated for 30 minutes at 80-85°C. After heating, ethanol was added to replace evaporated solvent. Thesupernatant and solids were separated by centrifugation followed byfiltration. The supernatant was analyzed by HPLC to determine the NAGcontent using standard methods, including a BioRad HPX-87H column heatedto 60° C., 0.01 N H₂SO₄ mobile phase at 0.6 mL/minute, and a refractiveindex detector.

Suitable blanks (samples with no added NAG) of each food type wereanalyzed to assess interferences. Mass changes between dry mixes andbaked final product were tracked to permit accurate recoverycalculations.

Using the AOAC method NAG was recovered as follows: For bread, the drybread mix and baked product yielded 93% and 80% recoveries of NAG,respectively. For cookies, the dry cookie mix and baked recoveries ofNAG were 78% and 68%, respectively. Therefore, the majority of NAG isunchanged when exposed to a high temperature, and is available to asubject upon ingestion of the heated food or beverage supplemented withNAG. These results compare favorably to the acid-extraction method ofrecovery of 100% and 77%, respectively.

EXAMPLE 16 Determination of the Amount of Glucosamine Present followingHeating

To determine the amount of glucosamine that remains following heating orpasteurization of food products and beverages, the following methodswere used. Food samples were homogenized prior to analysis. Wherepossible, samples were frozen and ground. Baked goods were crumbled andmixed to produce a homogeneous material.

A food or beverage sample as described in the preceding examplescontaining 5 to 20 mg of GLCN was dispersed in 25 g of 1.0 N HCl in a50-mL polypropylene centrifuge tube and capped tightly. The sample wasmixed for 30 seconds using a vortex mixer, then placed in a water bathat 37° C. The sample was removed from the water bath at 15-minuteintervals, mixed for 30 seconds on a vortex mixer, then returned to thewater bath. This cycle was repeated until the sample had been in thewater bath for one hour.

After heating, the sample was mixed for 30 seconds on a vortex mixer,then centrifuged for 10 minutes to separate the liquid and solid phases.Fats, oils or lipids in the sample formed a third layer at the top ofthe tube. A 1-g aliquot of the aqueous sample portion was diluted100-fold with deionized water, then transferred to an autosampler vialwith filter cap.

The free glucosamine in prepared samples was determined using highperformance anion-exchange chromatography with pulsed amperometricdetection (HPAEC-PAD). The system included an EG40 eluent generator,GP50 gradient pump, AS40 autosampler, LC25 column oven, and ED40electrochemical detector, all produced by Dionex Corporation (Sunnyvale,Calif.).

The method was adapted from Dionex Corporation Technical Note 40. ADionex CarboPac PA-20 column was used in place of the PA-10 described inthe Technical Note. The eluent was 8 mM KOH at 0.5 mL/min. The columnand detector were maintained at 30° C. The injection volume was 10 μL.The standard was glucosamine hydrochloride at 10.8 mg/L. Fermentationbroth samples were diluted five-fold with deionized water, ASTM Type II,and filtered through 0.2μ vial filters in the autosampler. Multiplestandards were analyzed before and after each sample set. The resultsare shown in Table 14. TABLE 14 Percent of NAG and GLCN recoveryfollowing heating. Sample Temp° F. Temp° C. Time NAG % GlcNHCl %Chocolate morsels (Example 4)  52 1 min 97 97 Rice Krispies ®(Example 1) 165  74 <10 min 106 Bread mix (Example 9) 250 121 50 min 10046 Sugar cookies (Example 2) 375 191 10 min 77 50 Corn muffin mix(Example 7) 400 204 18 min 77 13 Tea (Example 12) Room temp 98 96 Tea(Example 12)  90 20 sec 98 94 Tea (Example 12)  71 20 min 98 100 Tea(Example 12) boil 5 min 96 93

As shown in Table 14, there was some degradation of NAG and GLCN whenthe food products were baked or boiled. NAG was lesser affected byheating than was GLCN. The amount of NAG recovered ranged from about77%-100%, while the amount of recovery for GLCN varied more widely fromabout 13%-100%, for example about 46%-100%.

In view of the many possible embodiments to which the principles of thisdisclosure may be applied, it should be recognized that the illustratedembodiments are only particular examples of the disclosure and shouldnot be taken as a limitation on the scope of the disclosure. Rather, thescope of the disclosure is in accord with the following claims. Wetherefore claim all that comes within the scope and spirit of theseclaims.

1. A beverage comprising: a heat pasteurized NAG beverage, wherein thebeverage comprises at least about 0.01 g NAG per serving.
 2. Thebeverage of claim 1, wherein the heat pasteurized NAG beverage comprisesat least about 250 mg to about 1500 mg NAG per serving.
 3. The beverageof claim 1, wherein the heat pasteurized NAG beverage is at atemperature of at least about 160° F.
 4. The beverage of claim 3,wherein the heat pasteurized NAG beverage is at a temperature of atleast about 180° F.
 5. The beverage of claim 1, wherein the heatpasteurized NAG beverage is at a temperature of about 161° F. to about300° F.
 6. A method of preparing a beverage, comprising providing abeverage; adding at least about 0.01 g NAG per serving to the beverageto form a NAG beverage; and heat pasteurizing the NAG beverage at atemperature of least about 160° F.
 7. The method of claim 6, wherein theNAG beverage is heat-pasteurized at a temperature of at least about 200°F.
 8. The method of claim 6, wherein an amount of NAG present in the NAGbeverage is about 250 mg to about 1500 mg NAG per serving.
 9. The methodof claim 6, wherein the NAG is derived from fungal biomass containingchitin.
 10. A food product comprising: a NAG food product comprising atleast about 0.01 g NAG per serving, wherein the NAG food product is at atemperature of at least about 160° F.; and an absence of shellfishproteins.
 11. The food product of claim 10, wherein the NAG food productis at a temperature of at least about 200° F.
 12. The food product ofclaim 10, wherein the food product is a flour- or grain-based product.13. The food product of claim 1 0, wherein an amount of NAG present inthe NAG food product is about 250 mg to about 1500 mg NAG per serving.14. A method of preparing a food product, comprising providing a foodproduct; adding a first amount of NAG derived from fungal biomasscontaining chitin to the food product to form a NAG food product,wherein the NAG food product comprises at least about 0.01 g NAG perserving; and heating the NAG food product to a temperature of at leastabout 160° F.
 15. The method of claim 14, wherein the heating comprisesbaking, broiling, or boiling the NAG food product.
 16. The method ofclaim 14, wherein the first amount of NAG present in the NAG foodproduct is about 250 mg to about 1500 mg per serving.
 17. The method ofclaim 14 wherein the NAG food product is heated to a temperature of atleast about 200° F.
 18. The method of claim 6, wherein at least about0.007 g NAG per serving remains in the NAG beverage after heatpasteurizing.
 19. The method of claim 14, wherein a second amount of NAGpresent in the NAG food product after heating the NAG food product is atleast about 70% of the first amount of NAG present in the NAG foodproduct before heating the NAG food product.
 20. The beverage of claim 1or the food product of claim 10, wherein NAG comprises at least 1% of asweetener in the beverage or food product.
 21. A non-acidified foodproduct, comprising: a food product comprising at least about 0.01 g NAGper serving; thereby generating a NAG food product, wherein the NAG foodproduct is at a temperature of at least about 160° F.